BETA-GLUCURONIDASE AND RELATED GLYCLOSIDASES DEMONSTRATED BY INDOLYL SUBSTRATES IN LYMPHATIC TISSUE
Abstract
Histochemical and cytochemical techniques for detecting enzyme activity have been difficult to apply to the lymphatic system. Tissue glycosidases were demonstrated with techniques that show sharp intracellular localization and that are highly specific, although the heterogeneity and functional changes in component cells may still present interpretive difficulties. Halogen-substituted indolyl substrates were developed for beta- galactosidase, beta-glucosidase, beta-2-deoxyglucosidase, and beta-fucosidase. The most recently developed substrate for beta-glucuronidase is 5-bromo-4- chloroindol-3-yl-beta-D-glucopyruroniside. The final reaction product in all cases is identical: 5,5'-bromo-4,4'-chloroindigo. This intensely blue-green, finely granular precipitate is substantive and insoluble so that tissues can be dehydrated and permanently mounted. Specific inhibition for each substrate was demonstrated with analogue lactones. Differences in the distribution and intensity of the resultant straining in the various cells and tissues of the lymphoid system for these enzymes offer a unique opportunity to study the development of functional changes in reticular cells, including enzymatic induction and antigenic response.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 1968
- Accession Number
- AD0834754
Entities
People
- Alfred C. Standen
- Bjarne Pearson
- John R. Esterly